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PRODUCT INFORMATION Laurdan Item No. 19706 CAS Registry No.: 74515-25-6 Formal Name: 1-[6-(dimethylamino)-2-naphthalenyl]- O 1-dodecanone MF: C24H35NO FW: 353.5 Purity: ≥98% N UV/Vis.: λmax: 230, 250, 259, 284, 362 nm Supplied as: A crystalline solid Storage: -20°C Stability: ≥2 years Information represents the product specifications. Batch specific analytical results are provided on each certificate of analysis. Laboratory Procedures Laurdan is supplied as a crystalline solid. A stock solution may be made by dissolving the laurdan in the solvent of choice, which should be purged with an inert gas. Laurdan is soluble in the organic solvent chloroform at a concentration of approximately 10 mg/ml. Description Laurdan is a membrane-permeable fluorescent probe that displays spectral sensitivity to the phospholipid phase of the cell membrane to which it is bound.1 Quantitation of generalized polarization (GP) of laurdan can be used to identify phospholipid phase. When excited at 340 nm, GP values are 0.6 and -0.2 for gel phase and liquid crystalline phase, respectively. GP does not change with polar head group or pH (in the range 4-10); it changes only with phase state. Reference 1. Parasassi, T., De Stasio, G., Ravagnan, G., et al. Quantitation of lipid phases in phospholipid vesicles by the generalized polarization of Laurdan fluorescence. Biophys. J. 60(1), 179-189 (1991). WARNING CAYMAN CHEMICAL THIS PRODUCT IS FOR RESEARCH ONLY - NOT FOR HUMAN OR VETERINARY DIAGNOSTIC OR THERAPEUTIC USE. 1180 EAST ELLSWORTH RD SAFETY DATA ANN ARBOR, MI 48108 · USA This material should be considered hazardous until further information becomes available. Do not ingest, inhale, get in eyes, on skin, or on clothing. Wash thoroughly after handling. Before use, the user must review the complete Safety Data Sheet, which has been sent via email to your institution. PHONE: [800] 364-9897 WARRANTY AND LIMITATION OF REMEDY [734] 971-3335 Buyer agrees to purchase the material subject to Cayman’s Terms and Conditions. Complete Terms and Conditions including Warranty and Limitation of Liability information can be found on our website. FAX: [734] 971-3640 [email protected] Copyright Cayman Chemical Company, 10/03/2019 WWW.CAYMANCHEM.COM.

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Influence of Cholesterol on Phospholipid Bilayers Phase Domains As Detected by Laurdan Fluorescence
120 Biophysical Journal Volume 66 January 1994 120-132 Influence of Cholesterol on Phospholipid Bilayers Phase Domains as Detected by Laurdan Fluorescence Tiziana Parasassi,* Massimo Di Stefano,* Marianna Loiero,* Giampietro Ravagnan,* and Enrico Grattont *Istituto di Medicina Sperimentale, Consiglio Nazionale Ricerche, 00137 Rome, Italy, and tLaboratory for Fluorescence Dynamics, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA ABSTRACT Coexisting gel and liquid-crystalline phospholipid phase domains can be observed in synthetic phospholipid vesicles during the transition from one phase to the other and, in vesicles of mixed phospholipids, at intermediate temperatures between the transitions of the different phospholipids. The presence of cholesterol perturbs the dynamic properties of both phases to such an extent as to prevent the detection of coexisting phases. 6-Lauroyl-2-dimethylaminonaphthalene (Laurdan) fluorescence offers the unique advantage of well resolvable spectral parameters in the two phospholipid phases that can be used for the detection and quantitation of coexisting gel and liquid-crystalline domains. From Laurdan fluorescence excitation and emission spectra, the generalized polarization spectra and values were calculated. By the generalized polarization phos- pholipid phase domain coexistence can be detected, and each phase can be quantitated. In the same phospholipid vesicles where without cholesterol domain coexistence can be detected, above 15 mol % and, remarkably, at physiological cholesterol concentrations,
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Disruption of Membrane Cholesterol Organization Impairs the Concerted Activity of PIEZO1 Channel Clusters
bioRxiv preprint doi: https://doi.org/10.1101/604488; this version posted April 11, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Disruption of membrane cholesterol organization impairs the concerted activity of PIEZO1 channel clusters P. Ridone1,*, E. Pandzic2,*, M. Vassalli3, C. D. Cox1,5, A. Macmillan2, P.A. Gottlieb4 and B. Martinac1,5 1The Victor Chang Cardiac Research Institute, Lowy Packer Building, Darlinghurst, NSW 2010, Australia, 2Biomedical Imaging Facility (BMIF), Mark Wainwright Analytical Centre, Lowy Cancer Research Centre, The University of New South Wales, NSW, 2052, Australia , 3Institute of Biophysics, National Research Council, Genova, Italy , 4Physiology and Biophysics, State University of New York at Buffalo, Buffalo, NY, 14214, USA , 5St Vincent's Clinical School, University of New South Wales, Darlinghurst, NSW 2010, Australia *These authors have equally contributed to this work Corresponding author: Dr. Boris Martinac Victor Chang Cardiac Research Institute Lowy Packer Building Darlinghurst, NSW 2010, Australia E-mail [email protected] Key words: Mechanosensitive channel, Cholesterol, STORM, Patch-Clamp, Fluorescence Microscopy. SUMMARY: The essential mammalian mechanosensitive channel PIEZO1 organizes in the plasma membrane into nanometric clusters which depend on the integrity of cholesterol domains to rapidly detect applied force and especially inactivate syncronously, the most commonly altered feature of PIEZO1 in pathology. 1 bioRxiv preprint doi: https://doi.org/10.1101/604488; this version posted April 11, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved.
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New Insights on the Fluorescent Emission Spectra of Prodan and Laurdan
J Fluoresc (2015) 25:621–629 DOI 10.1007/s10895-015-1545-x ORIGINAL PAPER New Insights on the Fluorescent Emission Spectra of Prodan and Laurdan Cíntia C. Vequi-Suplicy & Kaline Coutinho & M. Teresa Lamy Received: 12 December 2014 /Accepted: 23 February 2015 /Published online: 10 March 2015 # Springer Science+Business Media New York 2015 Abstract Prodan and Laurdan are fluorescent probes largely Introduction used in biological systems. They were synthetized to be sen- sitive to the environment polarity, and their fluorescent emis- The fluorescent probes Prodan (2-dimethylamino-6- sion spectrum shifts around 120 nm, from cyclohexane to propionylnaphthalene) and Laurdan (2-dimethylamino-6- water. Although accepted that their emission spectrum is com- dodecanoylnaphthalene) have been widely used in biological posed by two emission bands, the origin of these two bands is relevant systems [1–7]. They were synthetized [1]tobesen- still a matter of discussion. Here we analyze the fluorescent sitive to the environment polarity, so their emission spectra spectra of Prodan and Laurdan in solvents of different polar- shifts about 120 nm from cyclohexane to water [1, 8, 9]. ities, both by decomposing the spectrum into two Gaussian Moreover, when inserted into membranes, their emission bands and by computing the Decay Associated Spectra spectra is extremely dependent on the lipid bilayer phase (DAS), the latter with time resolved fluorescence. Our data (gel or fluid), the maximum of the spectrum shifting around show that the intensity of the lower energy emission band of 50 nm from one phase to the other [4, 6, 10]. Prodan and Laurdan (attributed, in the literature, to the decay Although Prodan and Laurdan have been extensively used, of a solvent relaxed state) is higher in cyclohexane than in their structure and electronic distribution, in different solvents, water, showing a decrease as the polarity of the medium in- are still a matter of discussion [11–13].
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Biomimetic Curvature and Tension-Driven Membrane
1 Biomimetic curvature and tension-driven membrane 2 fusion induced by silica nanoparticles 3 4 Marcos Arribas Perez1 and Paul A. Beales1,2,* 5 6 1 Astbury Centre for Structural Molecular Biology and School of Chemistry, University of Leeds, 7 Leeds, LS2 9JT, UK. 8 2 Bragg Centre for Materials Research, University of Leeds, Leeds, LS2 9JT, UK. 9 10 * Correspondence: [email protected] 11 12 1 13 Abstract 14 Membrane fusion is a key process to develop new technologies in synthetic biology, where 15 artificial cells function as biomimetic chemical microreactors. Fusion events in living cells are 16 intricate phenomena that require the coordinate action of multicomponent protein complexes. 17 However, simpler synthetic tools to control membrane fusion in artificial cells are highly desirable. 18 Native membrane fusion machinery mediates fusion driving a delicate balance of membrane 19 curvature and tension between two closely apposed membranes. Here we show that silica 20 nanoparticles (SiO2 NPs) at a size close to the cross-over between tension-driven and curvature- 21 driven interaction regimes initiate efficient fusion of biomimetic model membranes. Fusion 22 efficiency and mechanisms are studied by Förster Resonance Energy Transfer (FRET) and 23 confocal fluorescence microscopy. SiO2 NPs induce a slight increase in lipid packing likely to 24 increase the lateral tension of the membrane. We observe a connection between membrane 25 tension and fusion efficiency. Finally, real-time confocal fluorescence microscopy reveals three 26 distinct mechanistic pathways for membrane fusion. SiO2 NPs show significant potential for 27 inclusion in the synthetic biology toolkit for membrane remodelling and fusion in artificial cells.
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Laurdan Fluorescence Lifetime Discriminates Cholesterol Content from Changes in Fluidity in Living Cell Membranes
View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector 1238 Biophysical Journal Volume 104 March 2013 1238–1247 Laurdan Fluorescence Lifetime Discriminates Cholesterol Content from Changes in Fluidity in Living Cell Membranes Ottavia Golfetto, Elizabeth Hinde, and Enrico Gratton* Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine, California ABSTRACT Detection of the fluorescent properties of Laurdan has been proven to be an efficient tool to investigate membrane packing and ordered lipid phases in model membranes and living cells. Traditionally the spectral shift of Laurdan’s emission from blue in the ordered lipid phase of the membrane (more rigid) toward green in the disordered lipid phase (more fluid) is quantified by the generalized polarization function. Here, we investigate the fluorescence lifetime of Laurdan at two different emission wavelengths and find that when the dipolar relaxation of Laurdan’s emission is spectrally isolated, analysis of the fluorescence decay can distinguish changes in membrane fluidity from changes in cholesterol content. Using the phasor representation to analyze changes in Laurdan’s fluorescence lifetime we obtain two different phasor trajectories for changes in polarity versus changes in cholesterol content. This gives us the ability to resolve in vivo membranes with different properties such as water content and cholesterol content and thus perform a more comprehensive analysis of cell membrane heterogeneity. We demon- strate this analysis in NIH3T3 cells using Laurdan as a biosensor to monitor changes in the membrane water content during cell migration. INTRODUCTION From the biological perspective, cell membranes influence that provide guidelines for understanding the complexity of many cell functions and are involved in most of the cellular cellular membranes.
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Laurdan Identifies Different Lipid Membranes in Eukaryotic Cells
UC Irvine UC Irvine Previously Published Works Title Laurdan identifies different lipid membranes in eukaryotic cells Permalink https://escholarship.org/uc/item/9m80z238 ISBN 9781482209891 Authors Gratton, E Digman, MA Publication Date 2014 DOI 10.1201/b17634 License https://creativecommons.org/licenses/by/4.0/ 4.0 Peer reviewed eScholarship.org Powered by the California Digital Library University of California Laurdan Identifies 13 Different Lipid Membranes in Eukaryotic Cells Enrico Gratton and Michelle A. Digman CONTENTS 13.1 Introduction ..................................................................................................283 13.1.1 Spectroscopic Properties of Laurdan................................................283 13.2 The Phasor Approach to Spectral and Lifetime Analysis ............................287 13.3 The Lifetime Phasor Transformation and Its Interpretation .........................289 13.4 Results of the Analysis of the Emission of Laurdan Using Spectral and Lifetime Phasors in GUVs Model Systems .................................................. 291 13.5 The Lifetime Phasor for Laurdan in GUVs ..................................................292 13.6 Live Cell Membrane Fluidity .......................................................................295 13.6.1 Spectral Phasors................................................................................295 13.6.2 Lifetime Phasors in Live 3T3 Cells ..................................................297 13.7 Conclusions and Further Considerations ......................................................299
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UC Irvine UC Irvine Previously Published Works Title Water dynamics in glycosphingolipid aggregates studied by LAURDAN fluorescence. Permalink https://escholarship.org/uc/item/2601m9d4 Journal Biophysical journal, 75(1) ISSN 0006-3495 Authors Bagatolli, LA Gratton, E Fidelio, GD Publication Date 1998-07-01 DOI 10.1016/s0006-3495(98)77517-4 License https://creativecommons.org/licenses/by/4.0/ 4.0 Peer reviewed eScholarship.org Powered by the California Digital Library University of California Biophysical Journal Volume 75 July 1998 331–341 331 Water Dynamics in Glycosphingolipid Aggregates Studied by LAURDAN Fluorescence L. A. Bagatolli,*# E. Gratton,# and G. D. Fidelio* *Departamento de Quı´mica Biolo´ gica-CIQUIBIC, Facultad de Ciencias Quı´micas, Universidad Nacional de Co´ rdoba, Argentina and #Laboratory for Fluorescence Dynamics, University of Illinois at Urbana-Champaign, Urbana, Illinois USA ABSTRACT We have characterized the fluorescence properties of 6-dodecanoyl-2-dimethylamine-naphthalene (LAURDAN) in pure interfaces formed by sphingomyelin and 10 chemically related glycosphingolipids (GSLs).1 The GSLs contain neutral and anionic carbohydrate residues in their oligosaccharide chain. These systems were studied at temperatures below, at, or above the main phase transition temperature of the pure lipid aggregates. The extent of solvent dipolar relaxation around the excited fluorescence probe in the GSLs series increases with the magnitude of the glycosphingolipid polar headgroup below the transition temperature. This conclusion is based on LAURDAN’s excitation generalized polarization (GPex) and fluores- cence lifetime values found in the different interfaces. A linear dependence between the LAURDAN GPex and the intermo- lecular spacing among the lipid molecules was found for both neutral and anionic lipids in the GSLs series.
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Naphthalene with Lipid Environments
Photochemistry and Photobiology, 1999, 70(4): 557-564 A Model for the Interaction of 6-Lauroyl-2-(N,Ak dimethy1amino)naphthalenewith Lipid Environments: Implications for Spectral Properties Luis A. Bagatolliin2*,Tiziana Parasassi3,Gerard0 D. Fidelio2and Enrico Gratton’ ’Laboratory for Fluorescence Dynamics, University of Illinois at Urbana-Champaign, Urbana, IL, USA; *Departamento de Quimica Biol6gica-CIQUIBIC, Facultad de Ciencias Quimicas, Universidad Nacional de C6rdoba, Argentina and 31stituto di Medicina Sperimentale, CNR, Roma, Italy Received 27 January 1999; accepted 24 June 1999 ABSTRACT to proteins and associated with lipids (1-3). This family of probes includes 6-propionyl-2-(N,N-dimethylamino) Although 6-lauroyl-2-(N,N-dimethylamino)naphthalene naphthalene (PRODAN),? 6-lauroyl-2-(N,N-dimethylamino) (LAURDAN) is now widely used a probe for lipid sys- as naphthalene (LAURDAN) and 2’-(N,N-dimethylamino)-6- tems, most studies focus on the effect of the lipid envi- naphthoyl-4-trans-cyclohexanoic acid (DANCA) (Fig. 1). ronment on its emission properties but not on the exci- These probes all possess large excited-state dipoles. In polar tation properties. The present study is intended to inves- solvents, and when the molecular dynamics of solvent di- tigate the excitation properties of LAURDAN in diverse poles occur on the time scale of the probe’s fluorescence lipid environments. To this end, the fluorescence prop- lifetime, a reorientation of solvent dipoles around the probe erties of LAURDAN were studied in synthetic ester and excited-state dipole may occur. The energy required for this ether phosphatidylcholines and sphingomyelin vesicles reorientation results in a red shift of the fluorescence emis- below, at and above the corresponding lipid main phase- sion spectrum.
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Alterations in the Properties of the Cell Membrane Due to Glycosphingolipid
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The Actin Homologue Mreb Organizes the Bacterial Cell Membrane
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